Title: | Cell wall trapping of autocrine peptides for human G-protein-coupled receptors on the yeast cell surface |
Author(s): | Ishii J; Yoshimoto N; Tatematsu K; Kuroda S; Ogino C; Fukuda H; Kondo A; |
Address: | "Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe, Japan" |
DOI: | 10.1371/journal.pone.0037136 |
ISSN/ISBN: | 1932-6203 (Electronic) 1932-6203 (Linking) |
Abstract: | "G-protein-coupled receptors (GPCRs) regulate a wide variety of physiological processes and are important pharmaceutical targets for drug discovery. Here, we describe a unique concept based on yeast cell-surface display technology to selectively track eligible peptides with agonistic activity for human GPCRs (Cell Wall Trapping of Autocrine Peptides (CWTrAP) strategy). In our strategy, individual recombinant yeast cells are able to report autocrine-positive activity for human GPCRs by expressing a candidate peptide fused to an anchoring motif. Following expression and activation, yeast cells trap autocrine peptides onto their cell walls. Because captured peptides are incapable of diffusion, they have no impact on surrounding yeast cells that express the target human GPCR and non-signaling peptides. Therefore, individual yeast cells can assemble the autonomous signaling complex and allow single-cell screening of a yeast population. Our strategy may be applied to identify eligible peptides with agonistic activity for target human GPCRs" |
Keywords: | "Autocrine Communication/*physiology Biological Assay/*methods Biotechnology/methods Blotting, Western Cell Wall/*metabolism Flow Cytometry Fluorescent Antibody Technique Humans Peptides/*isolation & purification/*metabolism Pheromones, Human/metabolism Pl;" |
Notes: | "MedlineIshii, Jun Yoshimoto, Nobuo Tatematsu, Kenji Kuroda, Shun'ichi Ogino, Chiaki Fukuda, Hideki Kondo, Akihiko eng Research Support, Non-U.S. Gov't Validation Study 2012/05/25 PLoS One. 2012; 7(5):e37136. doi: 10.1371/journal.pone.0037136. Epub 2012 May 18" |